(A) Percentages and numbers of Foxp3+CD4+CD8+ cells (DP Tregs) or Foxp3+CD8+ cells (CD8+ Tregs) inside the liver samples

(A) Percentages and numbers of Foxp3+CD4+CD8+ cells (DP Tregs) or Foxp3+CD8+ cells (CD8+ Tregs) inside the liver samples. H&E staining of liver slices, showing leukocyte adhesion to liver blood vessels (*, INF d7), perivascular infiltration (surrounded in black), and haemozoin deposition (green arrows). Black scale pub ?=?40 m. These results are representative of 3 repetitions. The inserts show hepatocytes from Control, INF d7, and INF d21 organizations, respectively.(TIF) pone.0081409.s002.tif (5.1M) GUID:?6286E557-FC4D-4AA2-82A9-A493BAA23232 Figure S3: Figures and phenotypes of liver leukocyte infiltrates after infection with iRBCs. Foxp3-GFP or CD11c-YFP mice were intraperitoneally infected or not with Pc-iRBCs. All animals were divided into three organizations: Control (non-infected; open columns); infected, on day time 7 of illness (INF d7; gray columns); or infected, on day time 21 of illness (INF d21; black columns). Liver cells were stained with MAbs against a panel of surface molecules to identify different types and subtypes of leukocytes. Total liver cell numbers were counted inside a haemocytometer chamber, and the final figures at each illness time were corrected according to the measured percentages. These results are representative of 3 repetitions.(TIF) pone.0081409.s003.tif (553K) GUID:?531A4FA9-A226-4601-8808-8BFDA16B51E4 Number S4: Illness with iRBCs and expression of cell surface molecules on liver Tregs and Tconv. Foxp3-GFP mice were intraperitoneally infected (or not) with Pc-iRBCs. All animals were divided into three organizations: Control (non-infected; open columns); infected, on day time 7 of illness (INF d7; gray columns); or infected, on day time 21 of illness (INF d21; black columns). Liver cells were stained with MAbs against a panel of surface molecules to identify different types and subtypes of leukocytes, as well as the manifestation levels of some proteins. (A) Percentages and numbers of Foxp3+CD4+CD8+ cells (DP Tregs) or Foxp3+CD8+ cells (CD8+ Tregs) inside the liver PI-1840 samples. (B) CD25 and CTLA-4 manifestation levels in CD4+ Tregs. (C) CD28, GITR, CD25, and CTLA-4 manifestation levels in CD4+ Tregs. These results are representative of 3 repetitions.(TIF) pone.0081409.s004.tif (769K) GUID:?B9FAA04B-9A54-4DF9-9890-AFC07B601ACF Number S5: Illness with iRBCs and expression of cell surface molecules about APCs inside the liver. PI-1840 CD11c-YFP mice were intraperitoneally infected or not with Pc-iRBCs All Rabbit polyclonal to AADACL3 animals were divided into three organizations: Control (non-infected; open columns); infected, on day time 7 of illness (INF d7; gray columns); or infected, on day time 21 of illness (INF d21; black columns). Liver cells were stained with MAbs against a panel of surface molecules to identify different types and subtypes of leukocytes as well as the manifestation levels of some proteins. (A) Percentages of plasmacytoid DCs (pDCs). (B) CD80 and CD86 expression levels in different subtypes of DCs. (C) Percentage of macrophages and CD80 expression levels. These results are representative of 3 repetitions.(TIF) pone.0081409.s005.tif (743K) GUID:?10EE9686-1738-4A1B-85E0-B3904BB67A86 Number S6: Illness with iRBCs prospects to the increased production of mRNA to iNOS and IL-10 inside the liver. Foxp3-GFP mice were intraperitoneally infected or not with Pc-iRBCs. All animals were divided into three organizations: Control (non-infected; open columns); infected, on day time 7 of illness (INF d7; gray columns); or infected, on day time 21 of illness (INF d21; black columns). Liver samples were frozen to further RT-PCR experiments. (A) iNOS mRNA levels. (B) IL-10 mRNA levels. These results are representative of three self-employed experiments.(TIF) pone.0081409.s006.tif (268K) GUID:?0713A5CB-A717-4A7B-A8E7-AF1F978C497F Video S1: mechanism of this accumulation and its potential immunological consequences. A massive liver build up of AS-iRBCs (Pc-iRBCs) was observed by intravital microscopy along with an over manifestation of ICAM-1 on day time 7 of the illness, as measured by qRT-PCR. Phenotypic changes were also observed in regulatory T cells (Tregs) and dendritic cells (DCs) that were isolated from infected livers, which show a functional part for Tregs in the rules of the liver inflammatory immune response. In fact, the suppressive function of liver-Tregs was tested, which demonstrated the capacity of these cells to suppress naive T cell activation to the same degree as that observed for spleen-Tregs. On the other hand, it is already known that CD4+ T cells isolated from spleens of protozoan parasite-infected mice are refractory to proliferate proliferative capacity in liver CD4+ T cells that were isolated on day time 7 of illness. It is also known that nitric oxide and IL-10 are partially involved in acute phase immunosuppression; we found high expression levels of IL-10 and iNOS mRNA in day time PI-1840 7-infected livers, which indicates a possible role.